Figure 5.

User-generated data can be uploaded and viewed in the context of complete PPI networks from public databases. (A) Vif network from GPS-Prot, including an uploaded dataset from AP-MS experiments (red-tagged nodes). Huwe1 is among several proteins in the uploaded dataset (Jager et al., submitted) that are not found in other databases (e.g., not present in Figure 2B), and were also previously identified by genetic/proteomic screens. (B) HIV Vif interacts with endogenous HUWE1 in 293 cells. 3xFLAG-tagged Vif, Vpr, and Nef were immunoprecipitated with anti-FLAG agarose beads. Lysates (L), remaining supernatant (S) and eluates (E) were analyzed by SDS-PAGE and Western blotting with antibodies as indicated. The same band is identified in the Vif pulldown by antibodies against the known CUL5 E3 ligase complex, anti-CUL5 (not shown) and anti-ELOB (TCEB2) as well as anti-Huwe1 antibodies, but not by the control anti-UPF1 antibody.